Effect of post-thaw dilution with autologous prostatic fluid on dog semen motility and sperm acrosome status

Abstract

The aim of this study was to evaluate the effect of post-thaw dilution with autologous prostatic fluid on motility parameters, longevity and acrosome status of frozen–thawed dog spermatozoa. After semen collection, seminal plasma was separated by centrifugation and stored frozen until use. Sperm pellets were diluted in two steps with an egg yolk–Tris extender to a final concentration of 5% glycerol and 0.5% Equex STM Paste. After thawing, semen was diluted 1:2 either with Tris buffer or with the autologous prostatic fluid. Motility was evaluated using a phase contrast microscope and a computer-assisted motility analyser system immediately after thawing and at hourly intervals up for 4 h at 38 °C. The status of acrosomes was assessed with Spermac ® stain at thawing and after 2 h of incubation. Motility and straight line velocity were initially higher in prostatic fluid-diluted samples (0 h and 0 and 1 h, respectively), but decreased to values similar to those of Tris-diluted samples in a time-dependent manner. In contrast, both the curvilinear velocity and amplitude of lateral head displacement were lower in prostatic fluid-diluted samples (1 and 3 h and 0, 1 and 3 h, respectively). The dilution did not have any significant effect on the percentage of acrosome-intact spermatozoa at either thawing or after 2 h. The pattern of motility of prostatic fluid-diluted samples suggests a reduction in hyperactivated motility with time, even though prostatic fluid neither prolonged spermatozoa longevity nor had any effect on the status of spermatozoa acrosomes.